孔雀石绿单克隆抗体的制备及其鉴定  

Preparation and identification of monoclonal antibody against malachite green

作　　者：龚朋飞[1] 王权[1] 陈永军[1] 

机构地区：[1]中国农业科学院上海兽医研究所兽医公共卫生实验室农业部动物寄生虫学重点实验室,上海200232

出　　处：《中国兽医科学》2007年第4期359-362,共4页Chinese Veterinary Science

基　　金：上海科技兴农重点攻关项目[沪农科攻字(2003)第9-10号]

摘　　要：用人工合成的无色孔雀石绿-兔血清白蛋白(LMG-RSA)作抗原免疫BALB/c小鼠,通过杂交瘤细胞融合技术筛选抗无色孔雀石绿的单抗,单抗经纯化后对其特异性和灵敏性进行了鉴定,探讨了建立孔雀石绿的免疫学检测方法。结果显示,经细胞融合和筛选得到了1株分泌抗无色孔雀石绿单克隆抗体的杂交瘤细胞5E9。经鉴定,该杂交瘤细胞的染色体数为98.82±5.6,间接ELISA检测小鼠腹水的抗体效价达1.3×105,该细胞连续培养10代以上,生物学性状稳定,间接竞争ELISA(ciELISA)结果显示,无色孔雀石绿50%抑制的质量浓度为18.5μg/L,除孔雀石绿外,与其他类似结构和相关类型的药物没有交叉反应,表明该单抗用于孔雀石绿的检测有较大的应用价值。A hybridoma cell line secreting monoclonal antibody against leuco-malachite green（LMG） was established after immunization of BALB/c mice with artificially synthesized LMG-rabbit serum albumin（LMG-RSA） as antigen. The stimulated splenocytes were fused with SP2/0 myeloma cells to produce hybridomas. The antibody was designated as 5E9. The number of chromosome of the hybridoma cell line was 98.82±5.6. The indirect ELISA titer of ascites was 1.3 × 10^5, and the 50% inhibitory concentration of detection of LMG was 18.5 μg/L as measured by the competitive inhibition enzyme-linked immunosorbent assay（ciELISA）. The McAb was quite specific as no cross-reactivity was observed with some drugs and with structurally related compounds. The McAb will be quite useful for the determination of MG residues.

关 键 词：孔雀石绿 单克隆抗体 间接竞争酶联免疫吸附试验 

分 类 号：S859.84[农业科学—临床兽医学] Q813.2[农业科学—兽医学]